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. 2006 Sep 15;36(2):138–146. doi: 10.1165/rcmb.2006-0253SM

Figure 5.

Figure 5.

NO indirectly regulates gelatinase activity at the epithelial wound edge. (A) In situ zymography analysis of gelatinase activity in wounded HBE1 cells. Confluent HBE1 cells were wounded and incubated as indicated in Materials and Methods in the absence (a), or presence of either scrambled siRNA (b), MMP-9 siRNA sequence (c), or the MMP activator p-CMB (d). In addition, untransfected cells (e and g) or MMP-9 siRNA transfected cells (f and h) were incubated in the presence of TNF-α (100 ng/ml) (e and f) or DETA NONOate (10 μM) (g and h). Arrows indicate direction of migration. (B) Recombinant human proMMP-9 was incubated with DETA NONOate (10 μM; X symbols), the nitrosating agent S-nitrosocysteine (200 μM; diamonds), or the MMP activator p-CMB (200 μM; circles), and activation was followed using fluorogenic MMP-9 substrate (DQ-Gelatin). Squares, control. (C) Analysis of uPA RNA in NHBE cells and protein expression in HBE1 cells after wounding and incubation in the absence or presence of DETA NONOate (10 μM). Left: representative RT-PCR analysis of uPA expression. Right: Analysis of uPA protein by immunohistochemistry. Arrows indicate direction of migration. Data represent mean ± SE from at least two separate experiments.